Investigation on oxidative stress of nitric oxide synthase interacting protein from Clonorchis sinensis

Abstract

Numerous evidences indicate that excretory-secretory products (ESPs) from liver flukes trigger the generation of free radicals that are associated with the initial pathophysiological responses in host cells. In this study, we first constructed a Clonorchis sinensis (C. sinensis, Cs)-infected BALB/c mouse model and examined relative results respectively at 3, 5, 7, and 9 weeks postinfection (p.i.). Quantitative reverse transcription (RT)-PCR indicated that the transcriptional level of both endothelial nitric oxide synthase (eNOS) and superoxide dismutase (SOD) gradually decreased with lastingness of infection, while the transcriptional level of inducible NOS (iNOS) significantly increased. The level of malondialdehyde (MDA) in sera of infected mouse significantly increased versus the healthy control group. These results showed that the liver of C. sinensis-infected mouse was in a state with elevated levels of oxidation stress. Previously, C. sinensis NOS interacting protein coding gene (named CsNOSIP) has been isolated and recombinant CsNOSIP (rCsNOSIP) has been expressed in Escherichia coli, which has been confirmed to be a component present in CsESPs and confirmed to play important roles in immune regulation of the host. In the present paper, we investigated the effects of rCsNOSIP on the lipopolysaccharide (LPS)-induced activated RAW264.7, a murine macrophage cell line. We found that endotoxin-free rCsNOSIP significantly promoted the levels of nitric oxide (NO) and reactive oxygen species (ROS) after pretreated with rCsNOSIP, while the level of SOD decreased. Furthermore, rCsNOSIP could also increase the level of lipid peroxidation MDA. Taken together, these results suggested that CsNOSIP was a key molecule which was involved in the production of nitric oxide (NO) and its reactive intermediates, and played an important role in oxidative stress during C. sinensis infection.

Role of Programmed Cell Death 4 (PDCD4)-Mediated Akt Signaling Pathway in Vascular Endothelial Cell Injury Caused by Lower-Extremity Ischemia-Reperfusion in Rats

“…Studies have confirmed that eNOS plays a protective role in vascular injury. It can regulate vasodilation, improve microcirculation, and reduce the occurrence of thrombus [20,21]. After examining the levels of PDCD4, Akt, and eNOS, and activation status of the Akt signaling pathway, we found that PDCD4 gene silencing activated the Akt signaling pathway and promoted the expression of eNOS.…”

TLR2 signal influences the iNOS/NO responses and worm development in C57BL/6J mice infected with Clonorchis sinensis

“…The amplified PCR products for wild-type and mutant TLR2 were 499 and 334 bp, respectively (Additional file 2 : Figure S1). Notably, the baseline levels of iNOS expression in the splenocytes of both mice types were remarkably higher than that in BALB/c mice [ 8 , 31 ]. The iNOS expression levels in the TLR2 wild-type and TLR2 mutant mice were remarkably induced by C. sinensis at 30 and 60 dpi, and the latter had higher iNOS expression level than the former in the corresponding period.…”

Current status and perspectives of Clonorchis sinensis and clonorchiasis: epidemiology, pathogenesis, omics, prevention and control

“…A study of the cancer-critical genes of C. sinensis -associated CCA showed that PSMD10 and CDK4 genes were upregulated, the tumour suppressor gene p53 and RB protein as well as BAX and caspase 9 were down-regulated, and PCNA was overexpressed in a C. sinensis- induced hamster CCA model [ 77 ]. Several researchers have proposed that oxidative stress might mediate liver fluke-associated carcinogenesis [ 78 – 80 ]. Serious pathological changes and increased DNA lesion products (resulting in the accumulation of lipid peroxidation products and the activation of COX-2 and 5-LOX) occurred in the hepatobiliary system of C. sinensis- infected mice, accompanied by the obvious activation of inducible NOS (iNOS) and malondialdehyde (MDA) [ 78 – 80 ].…”

“…Several researchers have proposed that oxidative stress might mediate liver fluke-associated carcinogenesis [ 78 – 80 ]. Serious pathological changes and increased DNA lesion products (resulting in the accumulation of lipid peroxidation products and the activation of COX-2 and 5-LOX) occurred in the hepatobiliary system of C. sinensis- infected mice, accompanied by the obvious activation of inducible NOS (iNOS) and malondialdehyde (MDA) [ 78 – 80 ]. The NOS-interacting protein of C. sinensis ( Cs NOSIP) might be a key participant in the oxidative stress observed in clonorchiasis [ 80 ].…”